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Biochemistry chapter 2

The document provides a comprehensive guide on SI prefixes and their corresponding units for length, volume, and mass, along with conversion methods between different units. It also details the preparation and storage of solutions, including weighing reagents, measuring volumes, and calculating molar and percentage solutions. Additionally, it covers the use of micropipettors and outlines tasks for group work on solution making in a laboratory setting.

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nickr2761
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3 views15 pages

Biochemistry chapter 2

The document provides a comprehensive guide on SI prefixes and their corresponding units for length, volume, and mass, along with conversion methods between different units. It also details the preparation and storage of solutions, including weighing reagents, measuring volumes, and calculating molar and percentage solutions. Additionally, it covers the use of micropipettors and outlines tasks for group work on solution making in a laboratory setting.

Uploaded by

nickr2761
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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Prefixes Commonly used SI units

Prefix Meaning Size relative Length Volume Mass


to
base unit

nano- billionth 10-9 nanometer (nm) nanoliter (nl) nanogram


(ng)

micro- millionth 10-6 micrometer (m) microliter (l) microgram


(g)

milli- thousandth 10-3 millimeter (mm) milliliter (ml) milligram


(mg)

centi- hundredth 10-2 centimeter(cm)

Base unit meter


liter gram

kilo- thousand 103 kilometer (km) kilogram (kg)

mega- million 106


giga- billion 109
Conversion:
Kilo- 103
Larger
base unit
Centi- 10-2
Milli- 10-3
Micro- 10-6
Smaller Nano- 10-9

Converting smaller to larger units:


divide (move decimal to left for the number
of exponents different)

Converting larger to smaller units:


multiply (move decimal to right for the number
of exponents different)
Preparation and storage of
solutions and pipetting
• Measuring
– Volumetric glassware: flasks, pipets and burets
(0.2% accuracy).
– Volumetric pipets: transfer, measuring, piston,
syringe
• Precision of results depends on precision of pipets
used.
– Reading meniscus in glassware: lowest point of
the liquid in the apparatus at eye level.
Preparation and storage of
solutions
• Weighing on analytical balance
– Be certain that balance is leveled.
– Clean balance.
– Tare or “zero” the balance and check for proper
operation.
– Tare balance with weigh boat in center of pan.
– Measure reagent to be weighed in weigh boat;
avoid air movement.
– When stable, record mass and finish weighing.
– Clean balance.
• We usually use distilled water or deionized water to
make solutions using these weighed reagents.
To make solution
• Start with volume of water about 0.5-0.75 of total
volume that you will need.
• Weigh out correct mass of reagent that you
need and add to beaker to mix with stir bar (Be
careful with stirring SDS (a detergent…why do
you think?)).
• Once dissolved, either use a volumetric flask of
desired volume or glass graduated cylinder to
bring up to correct final volume with water
(dH2O).
Calculating Solutions
1. Molar solutions:
1 mole of solute in 1 L of solution

mole: molecular weight in grams

1 M = molecular weight (g) of solute in 1 L


RULE: #grams = (moles/liter [M]) X (volume [L]) X (grams/mole [MW])

Example 1

What if you wanted to make 100 ml of 1 M Tris (MW: 121.1 g/M)?

# grams = (1M)(0.100)(121.1)
= 12.11 g

You would have to dissolve 12.11 g of Tris


in less than a final volume of 100 ml (50-75 ml) and
then bring up to final desired volume of 100 ml.
Best way to measure: 100 ml volumetric flask
Calculating Solutions
1. Molar solutions:
1 mole of solute in 1 L of solution
mole: molecular weight in grams

1 M = molecular weight (g) of solute in 1 L

#grams = (moles/liter [M]) X (volume [L]) X (grams/mole [MW])

Example 2
Make 500 ml of 5 M NaCl. (NaCl MW = 58.44)

# grams = (5 M)(0.5)(58.44)
= 146.1 g, dissolve in 350 ml, bring up to a
final volume of 500 ml
Best way to measure is in a 500 ml volumetric flask
Calculating Solutions

2. % Solutions:
weight of the solute per milliliter of solution

can be given as (w/v): meaning the solute is a “solid”


can be given as (v/v): meaning the solute is a “liquid”

w/v = # grams = (% grams/100) X (final volume, ml)

v/v = # ml = (% ml/100) X (final volume, ml)

*** % in the recipe is the FINAL percentage in the solution


Calculating Solutions
Examples:
2. % Solutions:

a. Make 30 ml of 5% BSA (w/v).

(w/v) – indicates that BSA is a “powder”

(5/100)*30 = 1.5 g of BSA in a final volume of 30 ml.

b. Make 10 ml of 0.5% Tween (v/v)

(0.5/100)*10 = 0.05 ml of Tween in a final volume of 10 ml.


Calculating Solutions
3. Dilutions from Concentrated Stock solutions

c1v1 = c2v2
Starting Final

C1: M of stock solution


V1:volume of stock solution, in liters need to add
C2:M wanted in final solution
V2: volume of final solution, in liters

*** make sure all units are the same on both sides of equation!
Calculating Solutions
3. Dilutions from Concentrated Stock solutions
Example 1:
You have a stock solution of 1 M Tris, and need to make 100 ml of 10 mM Tris.

C1: M wanted in final solution = 10 mM Tris


V1: volume of final solution, in liters = 100 ml
C2: M of stock solution = 1 M Tris
V2: volume of stock solution, in liters need to add = V

(0.01 M)(100 ml) = (1 M)(V) ( 0.01 M = 10 mM)

V = 1 ml of 1 M Tris added to 99 ml water


Calculating Solutions
3. Dilutions from Concentrated Stock solutions

Example 2:
You have a stock solution of 25X running buffer and you need to make
500 ml of 5X running buffer.

(5X)(500 ml) = (25X)(V)

V= 100 ml of 25X running buffer and 400 ml water


Calculating Solutions
4. Combination solutions: many buffers are made of a variety of
ingredients!

* calculate each component separately


* bring up to the final needed volume at the end!

Example: make 500 ml of 1X TBS. It is composed of:


10 mM Tris-HCl
150 mM NaCl
You have stock solutions of 1 M Tris and 5 M NaCl.

10 mM Tris:
(0.01 M)(500 ml) = (1 M)(v)
v= 5 ml of 1 M Tris

150 mM NaCl
(0.150 M)(500 ml) = (5 M)(v)
v = 15 ml

Final solution: 5 ml 1 M Tris, 15 ml 5 M NaCl, bring up to 500 ml with water


Use of Micropipettors
• Work by displacing air: creates a vacuum

• Always use the correct volume for the micropipette


• P10: 0.5 l or 1 l to 10 l
• P20: 1 l or 2 l to 20 l
• P200: 10 l or 20 l to 200 l
• P1000: 100 l or 100 l to 1000 l

• No OVERWINDING or UNDERWINDING: ruins calibration

• Use correct tips


• P10: clear tips, red box
• P20: yellow tips
• P200: yellow tips
• P1000: blue tips
Tasks
• Group Work on Solution making
• Follow directions on lab procedures
handout to carry out experiment:
• From protocol, Use of Micropipettors, Parts 1-3
• Make solution of Whey Protein of
choice according to Protocol for Week
2

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